LOCUS       sequence-337555-        6215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses INPP5K in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6215)
  AUTHORS   Feng Z, Yu CH
  TITLE     PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking
            and invasive cell migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 May 11;118(19). pii: 2017645118. doi:
            10.1073/pnas.2017645118.
  PUBMED    33947811
REFERENCE   2  (bases 1 to 6215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-05-11"; volume: "118(19). pii: 
            2017645118. doi"; pages: " 10.1073/pnas.2017645118"
COMMENT     Sequence Label: mCherry-INPP5K
FEATURES             Location/Qualifiers
     source          1..6215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      109..118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             115..822
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(260..278)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     528..547
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     722..745
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    835..859
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2204..2228)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2267..2401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2291..2310)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2345..2364
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2514..2534)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2514..2532)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2553..2569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2553..2569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2566..2588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2577..2593
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2601..2607,2608..2625,2626..2631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2646..2667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2784..2801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3035..3054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3714..4505,4506..4574))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4337..4356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4575..4679)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4705..5160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4842..4863)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5054..5073
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5231..6215
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggctagcgc caccatggtg
      121 agcaagggcg aggaggacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac
      181 atggagggct ccgtgaacgg ccgcgagttc gagatcgagg gcgagggcga gggccgcccc
      241 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
      301 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      361 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      421 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      481 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
      541 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
      601 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgccgaggtc
      661 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
      721 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
      781 gagggccgcc actccaccgg cggcatggac gagctgtaca agagatctcg aatcacaagt
      841 ttgtacaaaa aagttggcat gagctcgcgg aagctgagcg ggccgaaagg caggaggctc
      901 agcatacacg tcgtgacttg gaacgtggct tcggcagcgc cccctctaga tctcagtgac
      961 ctgcttcagc tgaacaaccg gaacctcaat cttgacatat atgttattgg tttgcaggaa
     1021 ttgaactctg ggatcataag cctcctttcc gatgctgcct ttaatgactc gtggagcagt
     1081 ttcctcatgg atgtgctttc ccctctgagc ttcatcaagg tctcccatgt ccgtatgcag
     1141 gggatcctct tactggtctt tgccaagtat cagcatttgc cctatatcca gattctgtct
     1201 actaaatcca cccccactgg cctgtttggg tactggggga acaaaggtgg agtcaacatc
     1261 tgcctgaagc tttatggcta ctatgtcagc atcatcaact gccacctgcc tccccacatt
     1321 tccaacaatt accagcggct ggagcacttt gaccggatcc tggagatgca gaattgtgag
     1381 gggcgagaca tcccaaacat cctggaccac gacctcatta tctggtttgg agacatgaac
     1441 tttcggatcg aggactttgg gttgcacttt gttcgggaat ccattaaaaa tcggtgctac
     1501 ggtggcctgt gggagaagga ccagctcagc attgccaaga aacatgaccc gctgctccgg
     1561 gagttccagg agggccgcct actcttcccg cccacctaca agtttgatag gaactccaac
     1621 gactatgaca ccagtgagaa aaaacgcaag cctgcatgga ccgatcgcat cctgtggagg
     1681 ctgaagcggc agccctgtgc tggccccgac actcccatac cgccggcgtc acacttctcc
     1741 ttgtctctga ggggctacag cagccacatg acgtacggca tcagcgacca caagcctgtc
     1801 tccggcacgt tcgacttgga gctgaagcca ttggtgtctg ctccgctgat cgtcctgatg
     1861 cccgaggacc tgtggaccgt ggaaaatgac atgatggtca gctactcttc aacctcggac
     1921 ttccccagca gcccgtggga ctggattgga ctgtacaagg tggggctgcg ggacgttaat
     1981 gactacgtgt cctatgcctg ggtcggggac agcaaggtct cctgcagcga caacctgaac
     2041 caggtttaca tcgacatcag caatatccct accactgaag atgagtttct cctctgttac
     2101 tacagcaaca gtctgcgttc tgtggtgggg ataagcagac ccttccagat cccgcctggc
     2161 tccttgaggg aggacccact gggtgaagca cagccacaga tctgcccaac tttcttgtac
     2221 aaagtggtga tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     2281 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2341 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2401 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2461 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     2521 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2581 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     2641 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2761 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4621 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4681 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4741 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4801 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4861 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4921 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4981 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5041 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5101 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5161 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5221 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     5281 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     5341 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     5401 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     5461 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     5521 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     5581 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     5641 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     5701 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     5761 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     5821 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     5881 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     5941 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     6001 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
     6061 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     6121 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     6181 gggaggtcta tataagcaat gctcgtttag ggaac
//
